Mechanism of PP2A affecting ubiquitination pathway in spermatogenesis.

PurposeUbiquitination plays a crucial role in various diseases. This study aims to explore the potential ubiquitination related genes in IPF.MethodsThe gene microarray dataset GSE24206 was obtained from GEO database. Subsequently, through differential expression analysis and molecular signatures database, we obtained 1734 differentially expressed genes and 742 ubiquitination related genes. Through the venn diagram analysis, we obtained 53 differentially expressed ubiquitination related genes. Then, gene-ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, protein-protein interactions (PPI) and gene set enrichment analysis (GSEA) were applied for the differentially expressed ubiquitination related genes. Finally, the expression of CDC20 and ITCH in IPF patients and cells were validated by qPCR and western blot assay.ResultsA total of 53 differentially expressed ubiquitination related genes (36 up-regulated genes and 17 down-regulated genes) were identified between 17 IPF patients and 6 healthy controls. GO and KEGG enrichment analysis of ubiquitination related genes mainly involved in regulation of protein ubiquitination, regulation of post-translational protein modification and ubiquitin mediated proteolysis. The PPI results demonstrated that these ubiquitination related genes interacted with each other. The GSEA analysis results for some of the hub genes mainly involved epithelial mesenchymal transition, inflammatory response, hypoxia, and apoptosis. The experiment expression level of CDC20 and ITCH in IPF patients and IPF cells were consistent with the bioinformatics analysis results.ConclusionWe identified 53 potential ubiquitination related genes of IPF through bioinformatics analysis. CDC20 and ITCH and other ubiquitination related genes may influence the development of IPF through epithelial mesenchymal transition and inflammatory response. Our research findings provide insights into the mechanisms of fibrosis and may provide evidence for potential therapeutic targets for fibrosis.

Study question How does GSK3α influence spermatogenesis, particularly in regulating sperm differentiation, morphology, and motility, and what molecular mechanisms underlie these effects? Summary answer We investigated GSK3α’s role in spermatogenesis using Gsk3α knockout mice to determine when morphological defects and motility impairments arise and uncover the underlying molecular mechanisms. What is known already Defects in spermatogenesis are a significant cause of male infertility. Around 2000 genes are involved in the regulation of spermatogenesis, with glycogen synthase kinase 3 (Gsk3) being one of them. GSK3α, an isoform of this serine-threonine kinase, plays an indispensable role in male fertility by regulating sperm function, and bioenergetics.Researchers had previously demonstrated that Gsk3α knockout (KO) mice exhibit reduced sperm counts, abnormal morphology, and impaired motility, leading to infertility. In contrast, males with only one mutated Gsk3α allele and male germ cell-specific Gsk3β knockout mice maintain normal fertility. Study design, size, duration The study employed a knockout mouse model (Gsk3α KO) to investigate the role of GSK3α in spermatogenesis. It utilized a comparative experimental design, analyzing wild-type (WT) and Gsk3α KO mice to identify developmental stages where sperm abnormalities emerge. The study incorporated molecular, cellular, and biochemical approaches Participants/materials, setting, methods Using real-time PCR and proteomic & Western blot analyses alongside cytochemical studies, and histological analysis we explored the influence of GSK3α on gene expression and protein phosphorylation during different stages of mouse sperm development. We evaluated its impact on both germ and somatic cells throughout the process. Main results and the role of chance Cell counts were significantly reduced in testicular, rete testicular, and caput epididymal spermatozoa of Gsk3α knockout (KO) mice. Morphological abnormalities, including bent heads, mid-pieces, and tails, were frequently observed during the transition from rete testicular to caput spermatozoa, suggesting defects in sperm differentiation. To validate these abnormalities, gene expression analysis was performed for Tnp2, a gene critical for sperm head development. Tnp2 expression was significantly lower in fully differentiated rete testicular spermatozoa of KO mice compared to wild-type (WT), whereas no difference was observed at the spermatid stage. Similarly, Prm1 (protamine 1), a marker of fully differentiated spermatozoa, showed significantly reduced expression in KO rete testicular spermatozoa relative to WT. Furthermore, phospho-proteomic analyses revealed differentially phosphorylated proteins associated with sperm maturation and differentiation, indicating disrupted signaling pathways essential for sperm development. Histological examination of testicular sections from WT and KO mice revealed a significant disruption in the transition from round spermatids to fully differentiated spermatozoa, further supporting the role of GSK3α in late-stage spermatogenesis. Collectively, these findings highlight the critical function of GSK3α in sperm differentiation, morphology, and maturation, ultimately impacting male fertility Limitations, reasons for caution This study necessitates the validation of phosphoproteomic results using Western blotting to confirm phosphorylation events. Future investigations should include transcriptomic analysis to explore gene expression changes. Additionally, examining isoform-specific expression of GSK3 in testicular somatic cells will provide further insights into its precise role in spermatogenesis and male fertility. Wider implications of the findings This study enhances understanding of GSK3α’s role in spermatogenesis, offering insights into male fertility regulation. Its findings could pave the way for novel treatments for infertility caused by spermatogenic defects and contribute to broader research on cellular differentiation and developmental biology, with potential applications in regenerative medicine Trial registration number No

Comparative transcriptomics of whole blood can be used to evaluate the systemic host response and its concordance between human and mouse malaria and aid the selection of appropriate models for translational malaria research.

PurposeUbiquitination is one of the important epigenetic modifications, influencing the development of various diseases. The objective of this study is to investigate the ubiquitination related genes in chronic obstructive pulmonary disease (COPD).MethodsThe gene microarray dataset from COPD patients and ubiquitination related genes were analyzed. Venn diagram analysis was used to intersect differentially expressed genes and ubiquitination related genes. The functional enrichment analysis of Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Set Enrichment Analysis (GSEA) were performed on differentially expressed ubiquitination related genes. Finally, we confirmed the expression of hub genes through qPCR and western blot experiments in clinical COPD patients and cell lines.ResultsWe identified 2,932 differentially expressed genes and 96 differentially expressed ubiquitination related genes. GO analysis indicated that the differentially expressed ubiquitination related genes were mainly enriched in post-translational protein modification and ubiquitin ligase complex. KEGG analysis showed that ubiquitination related genes were mainly involved in ubiquitin mediated proteolysis and TNF signaling pathway. GSEA analysis suggested that some hub genes are involved in allograft rejection, IL6/JAK/STAT3 signaling and inflammatory response. Our qPCR and western blot experimental results indicate that the expression of USP15 and CUL2 is higher in COPD group compared to the control group, consistent with the bioinformatics analysis.ConclusionOur bioinformatics analysis and experimental results suggest that USP15 and CUL2 may contribute to the progression of COPD through ubiquitination modification. To our knowledge, this is the first study to demonstrate the involvement of USP15 and CUL2 in COPD. Our results may provide new insights into the diagnosis and treatment of COPD.

Premenopausal women have blood pressure that is ~10mmHg lower than age-matched men. A study from our lab found that an olfactory receptor (OLFR558) is required for sex differences in blood pressure, as sex differences in blood pressure are entirely absent in Olfr558 knockout mice (Xu et al Science Advances 2024). However, there is a gap in our knowledge regarding how OLFR558 influences blood pressure, and, in the mechanism underlying the blood pressure differences between the sexes. We hypothesize that sex-specific differential protein expression between Olfr558 wild-type and knockout mice will yield novel insights into mechanisms driving sex differences in blood pressure. To pursue this, we collected kidneys from 8–9-week-old Olfr558 wild-type and knockout mice of both sexes for unbiased whole kidney proteomics (C57Bl6J; n=8 per genotype/sex). Whole kidney protein lysates were generated and sent to the Mass Spectrometry and Proteomics Core at JHU for TMTpro 16plex analysis (two plates; n=16 per plate). After filtering for proteins with an isolation interference of <30%, one protein group, and excluding spectra with at least one missing value and/or missing protein information, we found that the two plates shared 7,750 proteins. In addition, Plate 1 had 998 and Plate 2 had 972 unique proteins. To gain insight into why wild-type but not knockout mice display sex differences in blood pressure, we examined sex differences in protein expression. We first compared Olfr558 wild-type kidney samples (n=8 female vs n=8 males) and found 1,060 proteins which were differentially expressed by sex (p-value<0.05; log 2 fold change +/- 0.26). Of these 1,060 proteins which are differentially regulated by sex in wild-type mice, 865 were similarly differentially expressed by sex in knockout mice. However, there are 195 proteins that are differentially regulated by sex in wild-type but not knockout mice (female/male: 113 upregulated proteins, and 82 downregulated proteins). Preliminary pathway analysis indicates that a subset of these 195 proteins belong to endoplasmic reticulum proteins, some of which are associated with lipids. Conversely, we found that knockout mice have 776 proteins which are differentially expressed by sex in knockout but not wild-type kidneys (female/male: 501 upregulated proteins and 275 downregulated proteins). Based on pathway analysis, knockout mice have unique expression changes in proteins that correlate with cellular components including the ER membrane complex, cytoplasmic ribosomal proteins, mitochondrial respiratory chain complex III, and oligosaccharyltransferase complex, and, biological processes including protein insertion into ER membrane by stop-transfer membrane-anchor sequence. Better understanding of the relationship between sex hormones, blood pressure, and OLFR558 is pertinent to how physicians treat elevated blood pressure in men and women. JLP is supported by the American Heart Association Established Investigator Award, R21AG081683, R01DK139021, and R01DK137762. ADM is supported by NIH PREP grant R25GM109441 This abstract was presented at the American Physiology Summit 2025 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.

Axonemal dynein light intermediate polypeptide 1 is required for sperm formation and male fertility through association with MEIG1/PACRG complex in the manchette, involvement in a cargo transport system, intraflagellar transport, and sperm individualization.

BackgroundDifferential gene expression analysis using RNA-seq data is a popular approach for discovering specific regulation mechanisms under certain environmental settings. Both gene ontology (GO) and KEGG pathway enrichment analysis are major processes for investigating gene groups that participate in common biological responses or possess related functions. However, traditional approaches based on differentially expressed genes only detect a few significant GO terms and pathways, which are frequently insufficient to explain all-inclusive gene regulation mechanisms.MethodsTranscriptomes of survivin (birc5) gene knock-down experimental and wild-type control zebrafish embryos were sequenced and assembled, and a differential expression (DE) gene list was obtained for traditional functional enrichment analysis. In addition to including DE genes with significant fold-change levels, we considered additional associated genes near or overlapped with differentially expressed long noncoding RNAs (DE lncRNAs), which may directly or indirectly activate or inhibit target genes and play important roles in regulation networks. Both the original DE gene list and the additional DE lncRNA-associated genes were combined to perform a comprehensive overrepresentation analysis.ResultsIn this study, a total of 638 DE genes and 616 DE lncRNA-associated genes (lncGenes) were leveraged simultaneously in searching for significant GO terms and KEGG pathways. Compared to the traditional approach of only using a differential expression gene list, the proposed method of employing DE lncRNA-associated genes identified several additional important GO terms and KEGG pathways. In GO enrichment analysis, 60% more GO terms were obtained, and several neuron development functional terms were retrieved as complete annotations. We also observed that additional important pathways such as the FoxO and MAPK signaling pathways were retrieved, which were shown in previous reports to play important roles in apoptosis and neuron development functions regulated by the survivin gene.ConclusionsWe demonstrated that incorporating genes near or overlapped with DE lncRNAs into the DE gene list outperformed the traditional enrichment analysis method for effective biological functional interpretations. These hidden interactions between lncRNAs and target genes could facilitate more comprehensive analyses.

To study the differential gene expression and clinical significance in human immunodeficiency virus-infected individuals (HIVIIs) with penile squamous cell carcinoma. At our hospital from 2019 to 2020, we selected six samples of HIV-related penile squamous cell carcinoma for the experimental group and six samples of non-HIV-related penile squamous cell carcinoma for the control group. Transcriptome sequencing of sample mRNAs was performed by high-throughput sequencing. Differential gene expression analysis, differential Gene Ontology (GO) enrichment analysis and differential Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were carried out, and the reads per kilobase per million reads (RPKM) value was used as a measure of gene expression. A total of 2418 differentially expressed genes were obtained, of which 663 were upregulated and 1755 were downregulated (absolute value of logFC >1 and p value <.05). On the basis of the significance of the GO enrichment analysis, we found that the tumor protein p63 (TP63) gene was significantly upregulated and that the LIM domain only 4 (LMO4) gene was significantly downregulated in the experimental group compared with the control group. KEGG pathway analysis of the differentially expressed genes revealed that DNA replication was the most significant pathway associated with the upregulated genes and cell adhesion molecule (CAM) metabolism was the most significant pathway associated with the downregulated genes. The gene expression profiles of HIV-related penile squamous cell carcinoma and non-HIV-related penile squamous cell carcinoma are significantly different and involve significant GO enrichment and KEGG metabolic pathways, and this is very meaningful for the study of non-AIDS-defining cancers (NADCs). Differential expression of genes may be an important target for the prevention of penile squamous cell carcinoma in HIVIIs.

To investigate the differentiated whole genome expression profiling of gastric high- and low-grade intraepithelial neoplasia and early-stage adenocarcinoma. Gastric specimens from an upper magnifying chromoendoscopic targeted biopsy were collected from March 2010 to May 2013. Whole genome expression profiling was performed on 19 low-grade intraepithelial neoplasia (LGIN), 20 high-grade intraepithelial neoplasia (HGIN), 19 early-stage adenocarcinoma (EGC), and 19 chronic gastritis tissue samples using Agilent 4 × 44K Whole Human Genome microarrays. Differentially expressed genes between different types of lesions were identified using an unpaired t-test and corrected with the Benjamini and Hochberg false discovery rate algorithm. A gene ontology (GO) enrichment analysis was performed using the GeneSpring software GX 12.6. The differentially expressed gene was verified using a real-time TaqMan® PCR assay with independent tissue samples, including 26 LGIN, 15 HGIN, 14 EGC, and 20 chronic gastritis. The expression of G0S2 were further validated by immunohistochemical staining (IHC) in 24 LGIN, 40 HGIN, 30 EGC and 61 chronic gastritis specimens. The gene expression patterns of LGIN and HGIN tissues were distinct. There were 2521 significantly differentially expressed transcripts in HGIN, with 951 upregulated and 1570 downregulated. A GO enrichment analysis demonstrated that the most striking overexpressed transcripts in HGIN compared with LGIN were in the category of metabolism, defense response, and nuclear factor κB (NF-κB) cascade. While the vast majority of transcripts had barely altered expression in HGIN and EGC tissues, only 38 transcripts were upregulated in EGC. A GO enrichment analysis revealed that the alterations of the immune response were most prominent in the progression from HGIN to EGC. It is worth noting that, compared with LGIN, 289 transcripts were expressed at higher levels both in HGIN and EGC. A characteristic gene, G0/G1 switch 2 (G0S2) was one of the 289 transcripts and related to metabolism, the immune response, and the NF-κB cascade, and its expression was validated in independent samples through real-time TaqMan® PCR and immunohistochemical staining. In real-time PCR analysis, the expression of G0S2 was elevated both in HGIN and EGC compared with that in LGIN (P < 0.01 and P < 0.001, respectively). In IHC analysis, G0S2 immunoreactivity was detected in the cytoplasmic of neoplastic cells, but was undetectable in chronic gastritis cells. The G0S2 expression in HGIN was higher than that of LGIN (P = 0.012, χ (2) = 6.28) and EGC (P = 0.008, χ (2) = 6.94). A clear biological distinction between gastric high- and low-grade intraepithelial neoplasia was identified, and provides molecular evidence for clinical application.

Sperm Affects Head Sensory Neuron in Temperature Tolerance of Caenorhabditis elegans.

Cold coagulation and blood stasis (CCBS) syndrome is one of the common traditional Chinese medicine (TCM) syndromes of gynecological diseases. However, the molecular mechanism of CCBS syndrome is still unclear. Thus, there is a need to reveal the occurrence and regulation mechanism of CCBS syndrome, in order to provide a theoretical basis for the treatment of CCBS syndrome in gynecological diseases. The plasma proteins in primary dysmenorrhea (PD) patients with CCBS syndrome, endometriosis (EMS) patients with CCBS syndrome, and healthy women were screened using Label-free quantitative proteomics. Based on the TCM theory of "same TCM syndrome in different diseases," the differentially expressed proteins (DEPs) identified in each group were subjected to intersection mapping to obtain common DEPs in CCBS syndrome. The DEPs of gynecological CCBS syndrome in the intersection part were again cross-mapped with the DEPs of gynecological CCBS syndrome obtained by the research group according to the TCM theory of "different TCM syndromes in same disease" theory in the early stage, so as to obtain the DEPs of gynecological CCBS syndrome that were shared by the two parts. The common DEPs were subjected to bioinformatics analysis, and were verified by enzyme-linked immunosorbent assay (ELISA). A total of 67 common DEPs were identified in CCBS syndrome, of which 33 DEPs were upregulated and 34 DEPs were downregulated. The functional classification of DEPs involved in metabolic process, energy production and conversion, immune system process, antioxidant activity, response to stimulus, and biological adhesion. The subcellular location mainly located in the cytoplasm, nucleus, and extracellular. Gene ontology (GO) enrichment analysis showed that the upregulated DEPs mainly concentrated in lipid transport, cell migration, and inflammatory reaction, and the downregulated DEPs mostly related to cell junction, metabolism, and energy response. Protein domain enrichment analysis and clustering analysis revealed that the DEPs mainly related to cell proliferation and differentiation, cell morphology, metabolism, and immunity. The Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis clustering analysis showed that the upregulated DEPs were involved in inflammation and oxidative damage, while the downregulated DEPs were involved in inflammation, cell adhesion, cell apoptosis, and metabolism. The results of ELISA showed significantly increased levels of Cell surface glycoprotein MUC18 (MCAM) and Apolipoprotein C1 (APOC1), and significantly decreased levels of Vasodilator-stimulated phosphoprotein (VASP), Fatty acid-binding protein 5 (FABP5), and Vinculin (VCL) in patients with CCBS syndrome compared with healthy women. We speculated that cold evil may affect the immune process, inflammatory response, metabolic process, energy production and conversion, oxidative damage, endothelial cell dysfunction, and other differential proteins expression to cause CCBS syndrome in gynecological diseases.

Objective: To explore the difference in the expression profile of circular RNA in peripheral blood mononuclear cells between patients with mild and severe influenza pneumonia. Methods: From December 2018 to March 2019, 10 inpatients with mild and 10 inpatients with severe influenza pneumonia admitted to the Department of Infection and Clinical Microbiology of Beijing Chaoyang Hospital were included. Clariom™ D gene chip was used to explore the circRNA expression profiles of peripheral blood mononuclear cells (PBMC) isolated from the patients. The absolute value of the fold change (FC value)>2 and P<0.05 were used as the criteria to screen the differentially expressed circRNA, and the gene ontology (GO) enrichment analysis and the Kyoto Encyclopedia of Gene and Genome database (Kyoto Encyclopedia of Genes and Genomes, KEGG) signal pathway enrichment analysis were also performed. Results: The age of mild patients [M (P25, P75)] was 62.0 (34.5, 69.8) years old, including 4 males; the age of severe patients [M (P25, P75)] was 50.0 (37.0, 60.0) years old, all were males. A total of 137 differentially expressed circRNAs in PBMCs of mild and severe patients were screened. The numbers of up-regulated and down-regulated circRNAs in mild patients were 101 and 36, respectively. Among them, hsa_circ_0091073 (FC value=160.898, P<0.05) was the most significantly up-regulated circRNA and hsa_circ_0092219 (FC value =-17.630, P<0.05) was the most significantly down-regulated circRNA. GO enrichment analysis showed that a total of 111 secondary GO items were significantly associated with related differential expression of circRNA (P<0.05). The GO terms associated with upregulated circRNAs included DNA-templated transcription, regulation of DNA-templated transcription, regulation of transcription from RNA polymerase Ⅱ promoter, etc.; The GO terms associated with downregulated circRNAs included neutrophil degranulation, killing of cells of other organism, defense response to fungus, etc. KEGG signaling pathway analysis showed that there were 37 metabolic pathways related to differentially expressed circRNAs (P<0.05). Signaling pathways related to up-regulated circRNAs included nuclear factor-κB (NF-κB) signaling pathway, mitogen-activated protein kinase (MAPK) signaling pathway, tumor necrosis factor (TNF) signaling pathway, etc. Signaling pathways related to down-regulation of circRNAs included cancer transcription disorders, folate carbon pool, and other types of O-glycan biosynthesis. Conclusion: The expression of circRNA in PBMC of mild and severe influenza pneumonia patients is significantly different, and it may play a role in the pathogenic mechanism of influenza pneumonia through multiple signal pathways.

Acute respiratory distress syndrome (ARDS) is characterized by elevated pulmonary microvascular permeability; however, the role of circular RNAs (circRNAs) in this process remains unclear. Our study aims to discover the mechanism underlying the role of circRNA in pulmonary microvascular permeability in ARDS. We developed an in vitro model of ARDS using cultured human pulmonary microvascular endothelial cells (HPMECs) and lipopolysaccharide challenge. Genome sequencing revealed significant differences among the cells in the expression of circRNA. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses of the target genes were conducted. A circRNA-microRNA (miRNA)-messenger RNA (mRNA) competitive endogenous RNA (ceRNA) network was constructed. The GO enrichment analysis of the target genes in the ceRNA network was analyzed. The genome sequencing results identified 379 significantly upregulated circRNAs and 448 significantly downregulated circRNAs. The 10 circRNAs with the greatest degree of upregulation and the 10 circRNAs with the greatest degree of downregulation were identified. The GO enrichment analysis results indicated that differential circRNA expression may mediate the cellular response to DNA damage, including DNA repair. The KEGG analysis results indicated that the mechanism by which differential circRNA expression exerts these effects may involve the mitogen-activated protein kinases (MAPK) signaling pathway. The GO enrichment analysis of the target genes in the ceRNA network showed that the circRNAs were mainly involved in the fluid shear stress response, angiogenesis regulation, vascular development, and cell adhesion. The differential expression of circRNAs may play an important role in ARDS, especially in the control of HPMEC permeability. The circRNAs that were shown to have differential expression in response to vascular development and shear stress response could be used as biomarkers for the early prediction of ARDS disease and potential future therapeutic targets.

Zinc finger transcription factors play significant roles in the growth and development of plant and animal, but their function remains obscure in fungi. Lenzites gibbosa mycelia were extracted and sequenced by transcriptome analysis after growing on sawdust at different times to support mycelial growth of L. gibbosa in a nutrient matrix. Data bases used for analysis were the Kyoto encyclopedia of genes and genomes (KEGG) annotation, the cluster of orthologous groups of proteins (COG) and gene ontology (GO) annotation. Zinc finger class genes related to the growth and development of L. gibbosa were screened. GO annotation and enrichment analysis of differentially expressed genes were carried out. A total of 114.55 Gb Clean Data were obtained from the L. gibbosa transcriptome. The average Clean Data in each sample was 6.16 Gb. The relative efficiency of reads between each sample and the reference genome was 88.5% to 91.4%. The COG analysis showed that most zinc finger protein genes were related to replication, recombination and repair function. GO enrichment analysis showed that the expressed genes involved in cellular process, cell part and binding. We identified seventy-two expressed genes including seven up-regulated genes and sixty-five down-regulated genes by applying DESeq2 data analysis software. By comparing the significantly expressed genes with KEGG database, 66 annotated sequences were obtained, and 35 primary metabolic pathways were annotated. Pathway enrichment analysis showed that differentially expressed genes were significantly enriched in protein processing in endoplasmic reticulum and ubiquitin-mediated proteolysis pathways. Gene_11750 and gene_5266 are highly correlated with the growth and development of L. gibbosa and are closely related to protein processing in endoplasmic reticulum and ubiquitin-mediated proteolysis pathway. According to gene functional analysis, seven important differentially expressed genes related to the growth and development of L. gibbosa were identified.

Laryngeal cancer (LC) is a malignant tumor that occurs in the head and neck. Laryngeal cancer is one of the most common cancers of the neck and head, and its prognosis has always been poor. The incidence of LC increased gradually and showed an early rising trend. Laryngeal cancer is rarely studied in relation to immunity, Malignant tumors will change the state of the human body in various ways to adapt to their own survival and avoid the immune system. This study aims to explore the immune molecular mechanism of laryngeal cancer through bioinformatics analysis. The gene expression data was downloaded for 3 microarray datasets: GSE27020, GSE59102, and GSE51985. CIBERSORT algorithm was performed to evaluate immune cell infiltration in tissues between LC and healthy control (HC). Differentially expressed genes (DEGs) were screened. Functional correlation of DEGs were analyzed by Gene Ontology, Gene Set Enrichment Analysis and Kyoto encyclopedia of genes and genomes. Candidate biomarkers were identified by cytoHubba of Cytoscape. Spearman correlations between the above biomarkers and infiltrating immune cells were explored using R software analysis. The immune cell types of LC and HC were significantly different. Twenty-one DEGs were obtained by cross-screening. The function of DEGs is closely related to the number of immune cells. Five central genes (TNNT3, TNNI2, Desmin, matrix metallopeptidase 9 and cytotoxic T lymphocyte antigen 4) were screened. The HUB gene was demonstrated to have the ability to diagnose LC and HC with good specificity and sensitivity. The correlation between immune cells and biomarkers showed that hub gene was positively correlated with macrophages and dendritic cells, and negatively correlated with CD4 + T cell. TNNT3, TNNI2, Desmin, matrix metallopeptidase 9 and cytotoxic T lymphocyte antigen 4 can be used as diagnostic biomarker for LC. Macrophages, dendritic cells and CD4 + T cell may participate in the occurrence and development of LC.